Molecular cloning of Ruminococcus albus cellulase gene.

Cloning and expression in Escherichia coli of a cellulase gene from Ruminococcus flavefaciens.

An endoglucanase gene of Ruminococcus flavefaciens FD1 was cloned on the vector pEcoR251 to form the recombinant plasmid pMEB200. The cloned endoglucanase gene showed carboxymethylcellulase enzyme activity but no degradation of Avicel (FMC Corp., Philadelphia, Pa.) or filter paper. Carboxymethylcellulase activity was found during the late-exponential-growth phase and accumulated in the periplas...

Muralytic Activities of Ruminococcus albus 8.

Ruminococcus albus 8 was cultured with isolated alfalfa cell walls as the carbon source. The culture broth was assayed for muralytic enzyme activities. The effect, with respect to the production of such muralytic enzymes, of growing the microorganism on different carbon sources was also investigated. Also, the rates of solubilization and utilization by R. albus of individual alfalfa cell wall s...

Cellulose Adherence Factors in Ruminococcus albus

Preparation of the cellulase from the cellulolytic anaerobic rumen bacterium Ruminococcus albus and its release from the bacterial cell wall.

1. Most of the cellulase (CM-cellulase) elaborated by the rumen bacterium Ruminococcus albus strain SY3, which was isolated from a sheep, was cell-wall-bound. 2. The enzyme could be released readily by washing either with phosphate buffer or with water. 3. The amount of enzyme released was affected by the pH and ionic strength of the phosphate buffer. 4. The cell-wall-bound enzyme was of very h...

Molecular Cloning and Mutagenesis of Rat Glucocerebrosidase Gene

Purpose: The aim of this study was cloning the Gba enzyme in pUCBM21 plasmid, and making frame mutation on it and sequencing it. Materials and methods: mRNA was extracted from mouse spleen and glucocerebrosidase cDNA was synthesized and amplified by PCR with specific primers. cDNA was cloned in pUCBM21 and analyzed by restriction enzymes. A fragment of its sequence was deleted using MscI restr...